Apocarboxypeptidase B-sepharose: a specific adsorbent for peptides.
Apocarboxypeptidase B-Sepharose was prepared by immobilization of porcine carboxypeptidase B, followed by treatment of the column with o-phenanthrolin. This column efficiently adsorbed Met-enkephalin-Arg-Arg (YGGFMRR) in an optimum pH range of 6.5-7.5. The adsorbed Met-enkephalin-Arg-Arg was eluted at pH 4.0 and confirmed to be unaltered. In the apocarboxypeptidase B-Sepharose chromatography, Met-enkephalin-Arg-Arg or dynorphin 1-13 (YGGFLRRIRPKLK), substrates of carboxypeptidase B, was separated from Met-enkephalin (YGGFM), dynorphin B 1-9 (YGGFLRRQF), and beta-neo-endorphin (YGGFLRKYP) which do not react with the immobilized enzyme.
Yasuhara
T
T
Meiji Institute of Health Science, Odawara, Japan.
Ohashi
A
A
D000595:Amino Acid Sequence; D001051:Apoenzymes / Q000378:metabolism*; D001059:Apoproteins / Q000378:metabolism*; D043424:Carboxypeptidase B; D002268:Carboxypeptidases / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods; D003488:Cyanogen Bromide; D004723:Endorphins / Q000302:isolation & purification / Q000378:metabolism; D004745:Enkephalins / Q000302:isolation & purification / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D010455:Peptides / Q000302:isolation & purification* / Q000378:metabolism; D012685:Sepharose
D001051:Apoenzymes; D001059:Apoproteins; D004723:Endorphins; D004745:Enkephalins; D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D010455:Peptides; D012685:Sepharose; D002268:Carboxypeptidases; D043424:Carboxypeptidase B; D003488:Cyanogen Bromide